Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: Cells were fixed, TRAP stained, and visualized by light microscopy. (A) Representative 100x images of female (top) and male (bottom) mouse bone marrow cells defined by CD11b surface expression driven to form osteoclasts (OCs) with M-CSF (25ng/mL) and RANKL (50 ng/mL). Female OCs (black arrow) were magnified 3x and inset (top). TRAP positive cells with three or more nuclei (OCs) from (B) female and (C) male mice were enumerated from three random images at days 3 (top) and 5 (bottom) of RANKL treatment. Osteoclast size from female (B, right) and male (C, right) was measured by pixels per osteoclast using Adobe PhotoShop CS. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05, ** P ≤0.01, *** P ≤0.001).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Staining, Light Microscopy, Expressing

Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: (A) Representative flow cytometry of bone marrow from male and female Mk2 +/+ and Mk2 -/- mice. (B) Enumeration of Gr-1 - CD11b lo cells as a percent of the hematopoietic stem cell population (HSC, top) and CD115+ as a percent of Gr-1 - CD11b lo cells (bottom). (C) Enumeration of CD11b surface expression in female (left) and male (right) mouse bone marrow cells. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Flow Cytometry, Expressing

Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: (A) Representative western blot of pre-osteoclasts from male mice primed with M-CSF (10 ng/mL) for two days and stimulated with RANKL (100 ng/mL) for the indicated minutes (min). Densiometric analysis of (B) p38 (left) and (C) p-p38 (right) as a percent of GAPDH. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Sexual Dimorphism in MAPK-Activated Protein Kinase-2 (MK2) Regulation of RANKL-Induced Osteoclastogenesis in Osteoclast Progenitor Subpopulations

doi: 10.1371/journal.pone.0125387

Figure Lengend Snippet: (A) Representative confocal images of male dOCP lo cells treated with M-CSF (control) or M-CSF and RANKL for 30 minutes. A 3x magnified portion of p-p38 was inset (middle,left). (B) Pearson’s r correlation coefficients for NFATc1 and DAPI colocalization. (C) Pearson’s r correlation coefficients for p-p38 and DAPI colocalization. (D) Pearson’s r correlation coefficients for NFATc1 and p-p38 colocalization. Data are expressed as means ± SE compared to Mk2 +/+ controls (* P ≤0.05).

Article Snippet: Membranes were incubated in Cell Signaling primary antibodies: p-MK2 (Thr334, 27B7, rabbit mAb), p-p38 (Thr180/Tyr182, rabbit polyclonal), p38 (rabbit polyclonal), and GAPDH (14C10, rabbit mAb) at a 1:1000 dilution in 5% BSA in TBS-T overnight at 4°C.

Techniques: Control

Journal: Experimental neurology

Article Title: Dephosphorylated rather than hyperphosphorylated Tau triggers a pro-inflammatory profile in microglia through the p38 MAPK pathway.

doi: 10.1016/j.expneurol.2018.08.007

Figure Lengend Snippet: Fig. 3. Tau has a direct effect on p38 activation. Representative western blot (A) and quantification of phosphorylated p38 (p-p38) (B) and phosphorylated MK2 (p- MK2) (C) in microglia cells after treatment with PBS and Tau with (+) or without (−) the p38 activity inhibitor SB203580 (SB). Note that when SB was present, p38 was not activated in the presence of Tau, as shown by an indirect measure through p-MK2 (A-C). An N = 3 independent experiments were performed. Bars show means ± S.E. ∗p ≤0.05, # p ≤0.05, ++ p ≤0.01, and ∗∗∗p ≤0.001.

Article Snippet: They were then incubated overnight at 4 °C with the following: p38 MAPK antibody (#9212 Cell Signaling 1:1000 dilution); phospho-p38 MAPK (Thr180/ Tyr182) antibody (#9211 Cell Signaling 1:1000 dilution); MK2 antibody (sc-393,609 SCBT 1:100 dilution); phospho-MK2 (Thr334) antibody (sc-293,140 SCBT 1:1000 dilution); or GAPDH antibody (ab8245 Abcam 1:1000 dilution).

Techniques: Activation Assay, Western Blot, Activity Assay